FIGURE 6.

Effect of IL‐18 on steroidogenic enzyme expression. A‐C, The expression of three genes involved in steroidogenesis (CYP17A1, CYP11A1 and LH‐R) was determined in TC after 72‐h culture in complemented serum‐free McCoy's 5A media in presence or absence of IL‐18 (1000 pg/mL). Total mRNA was extracted from TC and reverse transcribed, and qPCR was performed. The housekeeping gene β‐actin was used to normalize gene expression. D‐F, The expression of the steroidogenic enzyme proteins (P450c17, P450scc and LH‐R) was determined in TC after 72‐h culture in the presence or absence of IL‐18 (1000 pg/mL). Proteins were then extracted and separated using electrophoresis on 4%‐12% (w/v) sodium dodecyl sulphate polyacrylamide gels. After electrotransfer to nitrocellulose membranes, the proteins were probed with P450c17, P450scc and LH‐R and GAPDH antibodies. The results are expressed as mean ± standard error of the mean (SEM) of six independent cultures. *Indicates a tendency (P < .05)